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Renal localization and quantification of uranium in rodent exposed to uranyl nitrate by LA-ICP-MS
Nagore Grijalba Marijuan, Alexandre Legrand, Valerie Holler, Celine Bouvier Capely
To cite this version:
Nagore Grijalba Marijuan, Alexandre Legrand, Valerie Holler, Celine Bouvier Capely. Renal lo- calization and quantification of uranium in rodent exposed to uranyl nitrate by LA-ICP-MS. 15th International Conference on Laser Ablation, COLA, Sep 2019, KAHULUI, HAWAII, United States.
2019. �hal-02635607�
Faire avancer la sûreté nucléaire
Rénal localisation and quantification of uranium in rodent exposed to uranyl nitrate
Context
Nagore GRIJALBA, Alexandre LEGRAND, Valérie HOLLER, Céline BOUVIER-CAPELY
Institut de Radioprotection et de Sûreté Nucléaire, PSE-SANTE/SESANE/LRSI, 31 Av de la Division Leclerc BP 17, 92262 Fontenay-aux-Roses Cedex, France
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The overexposure to uranium results in rénal toxicity, which is derived from an excessive accumulation of the element in the kidney. This may induce nephrotoxicity characterized by reduced glomerular filtration, exerting its toxic effect by Chemical and radiological action (alpha emitter) (1-3). In addition, its tissue distribution is heterogeneous mostly accumulating and producing pathological lésions in the S2 and S3 segments of the proximal tubule located in the cortical zone (100-fold above mean rénal concentration) (4-6). Chronic exposure (occupational exposure) to uranium, is linked to its bioaccumulation in kidney and could be associated with rénal dysfunction, an increased risk of cancer mortality and kidney failure (7-9).
Rodents were exposed to varying concentration of uranium in their drinking water for a period of 9 months. Total uranium analysis was done by conventional liquid ICP- MS after acid digestion using one kidney from each pair. In parallel, the other kidney was employed for LA-ICP-MS analysis (10). In order to perform quantitative LA- ICPMS bio-imaging analysis, synthetic matrix-matched laboratory standards and a normalisation strategy based on internai standard spiked gélatine were developed.
Quantitative images of cryo-sections of rat kidney revealed heterogeneous distribution of uranium within the rénal tissue, being the cortical concentration up to 123- fold higher than the medullary concentration.
Experimental proce
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External calibration Internai calibration
Objective:
Obtain a concentration - signal linear response to extrapolate the signal obtained from the sample.
Objective:
An internai standard (IS) is used for the normalisation of the signal to compensate signal variation and instrumental drift.
Matrix-matched standard = spiked homogenates of analogous tissue
Tissue homogénisation
Division in aliquots (6 std + 3 QC) Addition of standard solutions
Concentration vérification
by ID-ICP-MS Cryo-cutting(-20°C, 16 pim)
Requirements of the IS:
1.15 should behave in a similar manner to the analyte
2.15 must be in similar concentration în samples and standards
3.15 must be homogeneously distributed in samples and standards Gélatine as IS substrate:
1. Mimics the texture of biological tissue 2. Ease handling and fast préparation
3. Number of analytes and their concentrations can be adapted to the needs
(<s sptteed geUitméH-
std/GZC/sampLe
Laser beam
-> To ICP-MS
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3.50E-03 3.00E-03
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y = 0,0058x + 8E-05
R2 = 0 9936 4 *
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Slide coating with IS (Tm) spiked gélatine (10% m/v)
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IS normalised U concentration
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LA-ICP-MS quantitative uranium bio-imaging in kidney samples
Step 1: Analytical protocol validation by QC samples analysis
Guideline on bioanalytica! method validation (EMA et FDA, 2018)
Quantification of Quality Control samples (QC)
Why are they necessary and how must they be analysed?
> A QC is a biological matrix with a known quantity of analyte that is used to monitor the performance of a bioanalytical method and to assess the integrity and validity of the results of study samples
> They should cover the expected study sample concentration range
> Total QC should be at least six in number (low-, mid- and high-QC, in duplicate)
> They must be analysed în triplicate
■ LA-ICP-MS ■ Liquid ICP-MS
Step 2: Analysis of real samples
CorteJc Sélection of 3 samples
C. Poisson, J. Stéfani et al. Free Radical Research, 48(10), 2014
Sample U contamination? Ablation zone Whole organ average conc.
(ICP-MS, ng g'1 2 3 4 5 6 7 8 9 10)
R15 NO Cortex 4
R77 YES Cortex/Medula 3256
R78 YES Cortex 6729
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The calculated concentrations of the QC should be within ± 15-20% of the nominal value
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Expected conc Obtained conc
ng g1 ng g1
(Verified by ID-ICPMS) (Calculated by LA-ICPMS)
QC 1 20 ± 1 19 ± 3 (15% RSD)
QC 2 1042 ±92 1420 ±160(11% RSD)
QC 3 9384±1792 91281991(10% RSD)
Average U concentration in ablated zone -11700 ng g'1
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Conclusions
In this work the feasibility of an internai standard doped gélatine was assessed for its use in quantitative bio- imaging of U in kidney tissue by laser ablation coupled to ICP-MS. The proposed analytical bio-imaging approach was successfully applied for quantification of uranium of in biological samples (rat kidney). Quantitative images of cryo-sections revealed heterogeneous distribution of uranium within the rénal tissue, being the cortical concentration up to 123-fold higher than the medullary concentration which is in great accordance with the studies found in the literature. For the author's knowledge, this is the first quantitative study using an internai standard other than C13 to precisely quantify and localise uranium in kidney by LA-ICP-MS.
Bibliography
Acknowledgments
The authors acknowledge funding from Orano for the postdoctoral research fellow comprised in the UKCAN project. We also would like to thank IRSN for the access to PATERSON Platform and for its excellent technical and Personal assistance. In addition, we thank IRSN PSE- SANTE/SESANE/LRTOX and Dr. Yann Gueguen for the access to the preserved tissue bank. Additionally, thanks to Dr Christophe Pécheyran (PAMAL Platform, UPPA) for ceding us the software for data treatment and image reconstruction (FOCAL).
1. S. Keith, O Faroon, N. Roney et al. Toxicological profile for Uranium (Agency for Toxic Substances and Disease Registry, US, 2013).
2. D.P. Haley, R.E. Bulger, D.C. Dobyan, The long-term effects of Uranyl Nitrate on the Structure and Function of Rat Kidney", Virchows Arch [Cell Pathol], 41,181,1982.
3. L. Vicente-Vicente, Y. Quiros, F. Perez-Barriocanal et al., "Nephrotoxicity of Uranium: Pathophysiological, Diagnostic and Therapeutic Perspectives", Toxicological Sciences, 118 (2), 324, 2010.
4. S. Homma-Takeda, T. Kokubo, Y. Terada et al., Uranium dynamics and developmental sensitivity in rat kidney", Journal of Applied Toxicology, 33, 685, 2013.
5. T. Konishi, S. Kodaira, Y. Itakura et al., "Imaging uranium distribution on rat kidney sections through détection of alpha tracks using CR-39 plastic nuclear track detector", Radiation Protection Dosimetry, 1, 2018.
6. S. Homma-Takeda, Y. Terada, A. Nakata et al., Elemental imaging of kidneys of adult rats exposed to uranium acetate", Nuclear Instruments and Methods in Physics Research B, 267, 2167, 2009.
7. L. Stammler, A. Uhl, B. Mayer et al., Rénal effects and carcinogenicity of occupational exposure to uranium: a meta-analysis", Nephron Extra, 6 (1), 1, 2016.
8. S.G. Qu, J. Gao et al., Low-dose ionizing radiation increases the mortality risk of solid cancer in nuclear industry workers: a meta-analysis", Molecular and Clinical Oncology, 8 (5), 1590, 2018.
9. A.P. Golden, E.D. Ellis, S.S. Cohen et al., Updated mortality analysis of the Mallinckrodt uranium Processing workers, 1942-2012", International Journal of Radiation Biology, 9 (1), 1, 2019.
10. C. Poisson, J. Stéfani, L. Manens et al., « Chronic uranium exposure dose-dependently induces glutathione in rats without any nephrotoxicity », Free Radical Research, 48(2), 1218, 2014.
15th International Conférence on Laser Ablation - September 8 > 13 2019 (Maui-Hawaii, USA) m Nagore Grijalba Marijuan nagore. grijalbamarîjuan(g)irsn.fr
